ArticleParasitology research2019
Evaluation of propidium monoazide-based qPCR to detect viable oocysts of Toxoplasma gondii.
Article in Parasitology research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Improving biocide evaluation using propidium monoazide (PMA) viability staining technique.Scientific reports · 2025Article
- Establishment of a Protocol for Viability qPCR in Dental Hard Tissues.Microorganisms · 2024Article
- Propidium monoazide PCR, a method to determine OsHV-1 undamaged capsids and to estimate virus Lethal Dose 50.Virus research · 2024Review
- AmSphere · 2024Article
- Practical application of PMA-qPCR assay for determination of viable cells of inter-species biofilm ofBiology methods & protocols · 2024Article
- Can qPCR replace the standard plaque assay in the ASTM F1671 to assess personal protective equipment barrier performance?International journal of clothing science & technology · 2024Article
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- Contamination of Soil, Water, Fresh Produce, and Bivalve Mollusks withMicroorganisms · 2022Review
- Detection ofFrontiers in veterinary science · 2022Article
- New Avenues to DesignFrontiers in immunology · 2022Article
- A review on toxoplasmosis in humans and animals from Egypt.Parasitology · 2020Review
- Determination of the viability of Toxoplasma gondii oocysts by PCR real-time after treatment with propidium monoazide.Revista do Instituto de Medicina Tropical de Sao Paulo · 2020Article
- Seroprevalence of Toxoplasma gondii infection in sheep in Inner Mongolia Province, China.Parasite (Paris, France) · 2020Article
- Toxoplasma gondii Oocyst Infectivity Assessed Using a Sporocyst-Based Cell Culture Assay Combined with Quantitative PCR for Environmental Applications.Applied and environmental microbiology · 2019Article
- Environmental transmission ofFood and waterborne parasitology · 2019Review
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Information on the viability of Toxoplasma gondii oocysts is crucial to establish the public health significance of this environmental transmission stage that can contaminate water and foods. Interest for molecular-based methods to assess viability is growing and the aim of our study was to assess, for the first time, a propidium monoazide (PMA)-qPCR approach to determine the viability of T. gondii oocysts. Untreated and heat-killed (99 °C, 5 min) oocysts were incubated with PMA, a photoreactive DNA binding dye, and analyzed by confocal microscopy and flow cytometry to characterize oocysts' dye permeability. Different PMA concentrations (50 to 150 μM), incubation temperatures (22, 37, and 45 °C), amplicon length, selected targeted gene, and dyes (PMA, PMAxx™) were evaluated to define optimal conditions to discriminate specifically viable oocysts by PMA-qPCR. In theory, PMA binding to DNA would inhibit PCR amplification in dead but not in viable oocysts. Incubation at 22 °C with 100 μM PMA coupled to qPCR targeting a 123-bp sequence of the 529-bp repeat element allowed the distinction between viable and heated oocysts. However, the reduction of viability following heating of oocysts at high temperature was slight and, contrarily to reverse transcriptase-qPCR, the qPCR signal was not totally suppressed in heated suspensions. Therefore, PMA-qPCR is able to assess the impact of heating on T. gondii oocysts' viability but underestimates the efficacy of this treatment. The relevance of this technique to evaluate the efficacy of other inactivation processes and assess exposure of humans to this pathogen requires further investigations.
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